[Abstract] [Objective] To prepare IgY antibody against the common motif of metalloproteinases from Agkistrodon acutus venom, and to study its effect on the activity of snake venom [Methods] By analyzing the common sequences of various metalloproteinases from Agkistrodon acutus venom, we designed and synthesized the highly conserved antigen peptides PT1 and PT2, which are closely related to the enzyme activity; Hens were immunized with conjugated compounds KLH-PT1 and KLH-PT2 to obtain IgY antibodies. The titer of IgY and the cross-reaction characteristics with Agkistrodon acutus and Agkistrodon acutus venom were studied by ELISA and Western blot; Finally, the neutralizing activity of IgY was evaluated by anti-subcutaneous hemorrhage test in mice [Results] The results of ELISA and Western blot showed that anti-KLH-PT1 and anti-Agkistrodon acutus IgY could cross react with Agkistrodon acutus and Agkistrodon acutus venom, and the latter was significantly stronger than the former; Anti-KLH-PT2 IgY has no obvious cross reaction with Agkistrodon acutus venom and Agkistrodon acutus venom in vitro, but has a strong neutralizing toxicity in the anti-bleeding experiment in vivo, and is significantly superior to the former two [Conclusion] The IgY antibody that can cross react with various blood circulating snake venoms was prepared by designing the common sequence antigen peptide of metalloproteinase, which laid the foundation for the preparation of specific, efficient, low toxicity and broad-spectrum anti-hemorrhagic snake venom antibody
Post time: Jan-12-2023